PT78196B - Leukotriene antagonists - Google Patents

Abstract

The compounds represented by the formula (I) wherein m is 0, 1 or 2; p is 9, 10. 11, 12 or 13; R, is hvdrogen, amino or R₂ is hydroxyl, amino, -NHCH₂CONH₂; and X is -CO₂H, or wherein n is 1 or 2, Y is hydrogen or hydroxyl or amino with the proviso that when m is 0, R₁ is hyrogen, or pharmaceutically acceptable salt thereof have been found to be leukotriene antagonists and useful in the treatment of diseases in which leukotrienes are a factor, such as asthma.

Description

DESCRIPTION OF THE INVENTION

The "slow reaction substance of anaphylaxis" (SRL-A) has been shown to be a very potent bronchoconstrictive substance that is released primarily by the mastoid and basophil cells in case of antigen challenge. SRL-A has been proposed as the primary mediator in human asthma. In addition to their pronounced effects on lung tissue, SRL-A also produces changes in skin permeability and may be involved in acute skin allergic reactions. SRL-A has also been shown to cause depression of ventricular contraction and potentiation of the cardiovascular effects of histamine.

The discovery of naturally occurring leukotrienes and their relationship with SRL-A has increased interest in SRL-A and other arachidonate metabolites. Mouse, rat, guinea pig and man derived SRL-A have been characterized as mixtures of leukotriene-C (LTC)), leukotriene-D ((LTD)) and leukotriene-E (LTE)) , whose structural formulas are represented below:

co ₂ h

X · * 01ο ^LTC 4 R · 1 = Cys-Gly ^LTD 4 R ' = Cys-Gly LTE ,. R ' = Cis

By antagonizing the effects of LTC3, LTD4 and LTE4 or other pharmacologically active mediators on the airway smooth muscle of the terminal organ, the compounds and pharmaceutical compositions of this invention are valuable in the treatment of diseases in which leukotrienes are a factor, such as asthma.

The compounds of this invention are represented by the general structural formula (I):

s (ch ₂ ) _b chcor ₂

^CH3 < ^CH 2> _Px

CH-X

(I)

c = c

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-5etn which is m is 0, 1 or 2; p is 9, 10, 11, 12 or 13; R3 is hydrogen, O ^x II

amino or -NHCCH3; R 1 δ hydroxyl, amino, -NHCH 2 CO 2 H, -NCH 2 CO 2 H, CH 2

-NHCHCOâ,,H, -NHCHCOâ,,H or -NHCHâ,,CONHâ,,; β X is -COOH, -CH ₂ OH, CH ₃ CH (CH ₃ ) ₂

-CHCH-OH or -CH (CH ₀₎ COR, wherein η δ 1 or 2, Y is hydrogen or hi, ζ, ζ n

Oh and

R 2 is hydroxy or amino, provided that when m is 0, R 2 is hydrogen, or a pharmaceutically acceptable salt thereof.

Exemplary of the compounds of formula (I) are those in which the alkyl group adjacent to the double bond contains twelve carbon atoms (ie, p and ll), and are represented by formula (II):

12 ^H 25

C? C

S (CH) CHCQR_ R '2 2'm

CH-X

\

H

(II)

wherein m, R 2, R 3 and X are as described above.

Compounds of formula (II) wherein X is -CONH are derived from 3 (Z) -hexadecenoic acid, exemplified by 2- [2- (carboxyethyl) thio] -3 (Z) -hexadecenoic acid.

The compounds of formula (II) wherein X is -CH ₂ 0H are derivatives of 3 (Z) -hexadecen-l-ol, exemplified by 2 - / ""_{(2-carboxylic>} i ethyl) tio_7-3 (z) -hexadecen-1-ol.

Compounds of formula (II) wherein X is -CHHCHOH are deOH

of 4 (Z) -heptadecen-1,2-diol, exemplified by 3 - [(2-carboxyethyl) thio] -4 (Z) -heptadecene-1,2-diol.

Compounds of formula (I) wherein X is -CH (CHâ,,) COR,

of η, Y and R are as described above, are represented by formula (III):

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V

(CH3) CHCOR ^x 2m

(ΠΙ)

wherein m, η, ρ, Y, R 1 and R 2 are as described above.

Compounds of the formula (1) wherein η β 2 and p are 11 have a C 12 carbon structure and are designated 6 (z) -nonadecenoic acid derivatives. Also, when n is 1 and p is 11, the co-rapists.

The compounds of formula (III) have a carbon C. structure and are designated 5 (Z) -octadecenoic acid derivatives. Quanoo S n 2 and p is 9 the compounds of formula (lil) have a carbon backbone and C ^ ₇ acid derivatives are designated 6 (Z) -haptadecenôico. Fj. When n is 2 and p is 13, the compounds of the formula (II) have a carbon structure of Cj e and are designated 6 (Z) -hexanoic acid derivatives.

(6) -nonadecenoic acid derivatives of the compounds of the formula (1) wherein R 2 is hydroxy and R 2 is hydrogen;

two asymmetric centers, one of which is on the carbon atom 4 (ie the hydroxyl-substituted carbon atom), are

of the other on the carbon atom 5 (ie the carbon atom

substituted by thiol). This leads to the possibility of four this reoisomers for each compound. In practice, the compounds of this invention of the formula (1a) have been prepared in a mixture of the two stereoisomers, i.e. the 4R, 55β-isomer and the 4S, 5R isomer. The individual pure isomers are obtainable by preparative separation by high pressure liquid chromatography (HPLC) of the appropriate intermediate compounds, if such compounds have a third asymmetric center. In cases where a third asymmetric center is not available in the synthetic pathway of the desired compounds, one may introduce one employing an asymmetric protective group, such as an N-trifluoroethoxycarbonyl-i-prolyl ester. After separation of the individual stereoisomer, the protecting group is removed by standard procedures.

Qs derivatives of 6 (z) -nonadecenoic acid of formula (1)

are exemplified by the following compounds as the 4R,

62 368

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1

you

<f · "

-7-

55, the 4S, 5R isomer or mixture of the two isomers:

4-hydroxy-5 - [(2-carboxyethyl) thio] -6 (Z) -nonadecenoic acid;

acid 5 - / "(3-carboxymethylamino-3-oxopropyl) _thio> _7-4-hydroxy-6 (Z) -nonadecenóicoj

5 - [(2-amino-3-carboxymethylamino-3-oxopropyl) thio] -4-hydroxy-6 (z) -nonadecenoic acid

5- / ^- (3-carboxymethyl-N-methylamino-3-oxopropyl) -tio_7-4 Hydroxyiminopiperazine-6 (Z) -nonadecenoic acid}

5 - [(2-amino-3-carboxamidomethylamino-3-oxopropyl) thio] -4-hydroxy-6 (Z) -nonadeenoic acid}

4-hydroxy-5- [(carboxymethyl) thio] -6 (Z) -nonadecenoic acid}

5- [2- (aminocarbonyl) ethyl] thio] -4-hydroxy-6 (Z) -nonemonenoic acid;

4-hydroxy-5- (2-amino-2-carboxyethyl) thio-6- (Z) -nonadecenic acid; β

4-hydroxy-5 - [(3-carboxypropyl) thio] -6 (Z) -nonadecenoic acid.

The 5 (z) -octadecenoic acid derivatives of the compounds of formula (II) wherein Y is hydroxy also contain two Î ± centers. symmetric bonds at the carbon atom 3 and at the carbon atom 4, and therefore the possibility of four stereoisomers for each compound. to exist.

Specific compounds of formula (III) are the 5 (Z) -octanoic acid derivatives exemplified by the following compounds as a mixture of the four isomers:

3-hydroxy-4 - [(2-carboxyethyl) thio] -5 (z) octadecenoic acid.

Also representative of the compounds of formula (III): 4 - [(2-carboxyethyl) thio] -5 (z) -octadecenoic acid.

Compounds of formula (III) wherein Y is hydrogen are exemplified by 5 - [(2-carboxyethyl) thio] -6 (Z) -nonadecenoic acid.

The specific compounds of formula (III) are those derived from 6 (Z) -nonacenoic acid wherein R is amino and Y is hydroxy

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-8IS,

(3-carboxymethylamino-3-oxoprapyl) thio] -4-hydroxy-6 (Z) -nonadecenamide and 4-hydroxy-5 - [(2-carboxyethyl) thio] -6 (Z) - nonadecsnamide.

Additional compounds of the formula (III) are the (z) -heptadecenoic acid derivatives exemplified as a mixture of 4-hydroxy-5 - [(2-carboxyethyl) thio] -6 (Z) -heptadecenoic acid isomers.

Other compounds of the formula (III) are those derived from 6 (z) -heenoicosenoic acid in which R ó is hydroxy and X hidrox is hydroxy exemplified as a mixture of 4-hydroxy-5- [2- (2-carboxyethyl) thio] -6 (Z) -hexanoic acid.

The compounds of formula (I) wherein X is -CQ ₂ H, -CH? OH and -CHCH ₉ 0H are prepared according to the following synthetic routes,

OH

starting from 4-hydroxybut-2 (E) -ene-1-al-tetrahydropyranyl ether (1), wherein THP is a tetrahydropyranyl radical, as follows: Route A

OH = HCl = C

ch ₂ othp

(D

CH ₃ (CH ₂ ) _p CH = 1

(2)

CH ₃ (CH ₂ ) _p

(I.e.

c = tr h

Η H

^CH 2 ^0THR

->

CH ₂ OH

c ^h 3 (ch ₂ ) _Px

c = c H <

(3)

(4)

^C M ^CH 2>

THE

CH-CHCH2 OH

^R 1 Γ

HS (CH3) CHCOR,

01

R-,

11

S (CHâ,,) CHCOR

CH (CH) OH ₀ CHCHCH

ZZ

T = ^Z H OH C \

(5)

(6)

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S (CH ₂ ) _n CHCOR ₂

CHCHCH _O QH

Via B

R ·

71

^CH 3 ( ^CH 2)

C = C

OH

-9-

^g M ^ch 2> p.

x = c

CHCHO

H ^X H (6)

ch ₃ (ch ₂ ) _p

C = c _x Η H

H ^Z \

(7)

S (CHâ,,) CHCOR '' 2 '' 2

HCH "QH

(8)

Via C

CH3 _(CH2) ^

f ¹ '

( ^CH 2) _m CHC ° R ₂

CHCHO

->

^CH / ^{CH 2} V

c = c

l

^ _{(CH2) 2} · _fn CHC0R

CHCO H / z

(7) (9)

Compound (1) which is a known compound is reacted with the appropriate alkyltriphenylphosphonium ylide under Wittig conditions to give compound (3) hydrolyzed compound (3)

to give the compound (4) which is epoxidized to give the compound (5). The compound (5) is reacted with the mer. in which R ₂ 'and R ₂ ' are R ₂ and R ₂ respectively as described above or a moiety easily convertible to the desired sub-entity, such as an alkyl ester or trifluoromethylacetaraid, to give the compound (6) The compound (6) can be (a)

easily converted to the compounds of formula (I) wherein X is -CHCH2 OH or analogues thereof; or (b) divided oxidati

OH

varaent for the compound (7). The compound (7) may be (a) reduced

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and

-10-

for the compound (8), which is readily converted into the compounds of the formula (I) wherein X is -CF2 OH or the analogues thereof; or (b) is oxidized to the compound (9), which is readily converted into the compounds of the formula (1) wherein X is -CO-H ₂ or the analogues thereof.

Compounds of formula (III) wherein n is 2 and Y 6 hydrogen are readily prepared by making the appropriate thiol having the compound of formula (A)

Rt '

P

HS (CH ₂ ) _m CHCOR ₂ '(A)

where m is as described above and R ₂ 'and R ₂ ' are respectively R 1 and R ₂ or a radical easily convertible to the desired substituent, such as an alkyl or trifluoromethylacetamide ester, is reacted with a 4,5- (Z) -alkanoic acid of the formula (B)

0

/ \

CH ₃ (CH ₂₎ CH ₂ CHCH ₃ CH ₂ C0r

^P = c = ^{z z} H ^z H

where p is as described above, R 1 is R 2 or a radical which is readily convertible at the desired acid percents, such as COalk wherein alk is a lower alkyl group. The compounds of formula (A) containing thiol are known or readily prepared from a known compound using standard chemical transformations. The derivatives (B) of 4,5-epoxy-6 (z) -alkanoic acid are readily prepared from monooleyl succinate (10) according to the following synthetic route:

H0 ₂ CCH ₂ CH ₂ CO ₂ alk

(10)

0HCCH = CHCH ₂ CH ₂ CO ₂ alk

(13)

0

II

-> CHCCH ₂ CH ₂ CO ₂ ->

(11) (12)

THE.

-> OHCCH-bHCH ₂ CH ₂ CO ₂ alq ->

(14)

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-110

/ \

CH, (CH 3) CH-CHCH CHCOAl q

3 2/2 2 2

WH

(15)

The monomethyl succinate (10), wherein alkyl is methyl, was chlorinated with oxalyl chloride in dimethylformamide and methylene chloride to give 3-carbomethoxypropionyl chloride (11). The compound

(II) was catalytically hydrogenated over palladium-charcoal catalyst in the presence of 2,6-lutidine to give 3-carbomethoxypropionaldehyde (12) which was reacted with formylmethylene triphenylphosphorane under Wittig reaction conditions to give 5-carbethoxy- 2-pentenal (13). The compound (13) was epoxidized with

aqueous hydrogen peroxide in the presence of 1N sodium bicarbonate to give methyl 4,5-epoxy-6-oxohexanoate (14). The compound

(14) is reacted with alkyltriphenylphosphonium ylide to give

4,5-epoxy-6 (Z) -alkenate (15).

The 5 (Z) -alkanoic acid derivatives of formula (iii) wherein n is 1 and Y is hydroxy are prepared according to the synthetic route starting from 2- [2- (2-carbomethoxyethyl) thio] -3 (Z) -alkenyl (7) as follows:

I ¹

S (CHâ,,) CHCORâ μ2 Âμm 2

CH, (CH 2 CHCHO-

^{3 2} P /

_c. -e _x

Η H

-7 ch ₃ (ch ₂ )

S (CH ₂ ) _ro CHCOR ₂ '

£ ^ H HCH ₂ C0 ₂ alk -> (III ^X CC OH

z \

(7) (16)

Compound (7) was reacted with methyl acetate in the presence of lithium diisopropylamide to give methyl 3-hydroxy-4- (2-chloromethoxyethyl) thio] -5 (Z) -alkenenoate (16), which is saponified to give the compound of formula (III) wherein m is 1, R is hydrogen, R ₂ is hydrogen and R ₂ is hydroxyl.

The 6 (Z) -nonadecenoic acid derivatives of the formula (iii) wherein n is 2 and Y is hydrogen are prepared according to the route

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-12-

of 5-carboxyalkyl-pentanoic acid (17) as the sagas:

H0 ₂ CCH ₂ CH ₂ CH ₂ CH ₂ CO ₂ alk

(17)

( ¹ '

S (CH) CHCOR '

| 2 m 2

H0 ₂ CCH _C H ₂ CH ₂ CH ₂ C ₀₂ alk

(19)

H0 ₂ CCHCH ₂ CK ₂ CK ₂ C0 ₂ al _q

(18)

^R 1 '

I ¹

S (CHâ,,) CHCORâ € ¢ 2 m 2

- CHCHCH "CH _or CH _or CO" alq | 2 2 2 2

(20)

R. · 11

S (CH ₂ ) _m CHCOR ₂

^CH 3 ( ^CH 2)

CHCH ₂ CH ₂ CH ₂ CO ₂ alk

(III)

c = c

(21)

The 5-carbomethoxypentanoic acid (17) in methylalcohol was reacted with thionyl chloride and then with N-bromosuccinimide (NBS), followed by hydrolysis of the base to give 2-bromo-5-carbomethoxypentanoic acid (18 ). The compound (18) was made

Reaction with methyl 3-mercaptopropionate (m = 1, R 2 = H and R ₂ '= to OCH 2) yielding 5-carbomethoxy-2- (2-carbomethoxyethyl) thio] pentanoic acid (19). Compound (19) was first reacted with diborane and

dimethyl sulfoxide and trifluoroacetic anhydride to give 5-carbomethoxy-2- [2- (carbomethoxyethyl) thio] -pentanal (20),

compound (20) was reacted under Wittig conditions with alkyltriphenylphosphonium ylide to give methyl 5- [2- (carbomethoxyethyl) thio] -6 (z) -alkenate (21), which is saponified to give a compound of formula (iii) wherein m is 1, R _{2 is} hydrogen and R _{2 is} hydroxy.

The 5 (z) -octadecenoic acid derivatives of formula (iii) wherein n is 1 and X is hydrogen may be prepared according to the general synthetic route used for the preparation of the 6 (Z) -nonadecenoic acid derivatives, but starting with of 4-carboxyalkyl-butanoic acid.

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Compounds of formula (1) wherein R4 is amino may be prepared by initially forming the β-lactone of the appropriate compound of formula (1a) with trifluoroacetic anhydride and then reacting the lactone with ammonia to give the desired amide.

The present invention also encompasses the

4,5-epoxy-6 (Z) -alkanoic acid of the formula (B):

^c f ^{CH 2} V /

c = c

0

/ \ CH

CHCH ₂ CH ₂ CO ₃

(B)

in which p is 9, 10, 11, 12 or 13 and R 2 is R or a radical which is readily convertible to R 1, such that Q 1, wherein alk is an alkyl radical containing one to six carbon atoms.

In addition, this invention relates to (C):

includes the

compounds of the formula

S (CHâ,ƒ) CHCORâ, "jzmz

CH, (CH 3) CHCHO

2 z p. /

C = C

Η H

wherein m is 0, 1 or 2; p is 9, 10, 11, 12 or 13;

0 0

I read

-NHCF-1 or -NHCCH3, and R4 'is amino, -NHCH2COgR3,

(W)

NCH 2 CO 2 R 5,

-NH 2, HCO 2 R 3 ', -NHCHCO 2 R 3', -NHCH 2 CH 2 NH

CH 2 CH (CH ₂ ) ₃

alkyl having one to six atoms when m is 0, R 2 is hydrogen.

ch ₃

or OR4 'where R4' is a radi.

carbon dioxide, provided that

The compounds of the formulas (Β) θ (C) have the binding

double in the cis conformation. The structural similarities between these

compounds and the compounds of formula (I) contribute to the properties.

agonist ages of the leukotrienes of the compounds of formula (I).

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-14-

The leukotriene antagonist activity of the compounds of this invention is measured by the ability of the compound to inhibit leukotriene-induced contraction in guinea pig tracheal tissues in vitro and to inhibit leukotriene-induced bronchoconstriction in guinea pigs in vivo. The following methodologies were used:

In vitro: spiral tracheal strips of guinea pigs (adult males of the Hartley albino strain), about 2 to 3 mm wide in section and 3.5 cm in length, were plated in modified Krebs buffer in a 10 ml tissue bath coated and aerated continuously with 95% O ₂ /5% CO ₂ . The tissues were attached by silk suture to force shifting transducers for recording isometric tension. Tissues were equilibrated for 1 hour, pretreated for 15 minutes with meclofenamic acid (1æM) for removal of the intrinsic responses to prostaglandins, and then pretreated for a further 30 minutes with either the test compound or control-vehicle . A cumulative concentration response curve for the LTD2 in triplicate tissues was generated by successive increases in the concentration of the LTD4 bath. To minimize tissue-to-tissue variability, the contractions elicited by LTD4 were normalized as a percentage of the maximal response obtained for a reference agonist, carbachol (10%).

Calculations: The means of the triplicate response curves of the LTD4 concentration both in the presence and absence of the test compound were scored on plot paper. The concentration of LTD4 required to elicit 30% of the concentration elicited by carbachol was measured and defined as ECjq. The pA ₂ value for the test compound was determined by the following equations:

^ £ 30 (P ^res ® ^N and Po test compound)

1. - = dose ratio = X

EC _3Q (control-vehicle presence)

2. Κθ = test compound concentration / (XI)

3. pA ₂ s-log. Kg

62 368

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In vivo: anesthetized guinea pigs (adult males of the Hartley albino strain), breathing spontaneously, were controlled on a Buxco pulmonary mechanics computer. The variations of the resis. (R ^) were calculated by the computer on a respiration-to-respiration rate at isovolomic points from air-pressure and transpulmonary pressure measurements by differential pressure transducers. The animals received or test compound or control-vehicle intravenously into the jugular vein. The LTD4 was then injected into the jugular vein. Bronco-constriction resulted in percent changes in airway resistance relative to baseline values obtained prior to injection of the test compound or control-vehicle. Each guinea pig received either vehicle-control or test compound.

Calculations: The mean of 3 - 6 animals per treatment was calculated using percent changes of lung parameters for control animals and animals treated with test compound. The mean percent inhibition by the test compound was calculated from the following equation:

r _l r _l

(control vehicle) - (test compound)

- X ₁₀₀

^r l

(control-vehicle)

The compounds of this invention possess antagonistic biosignificant activity against leukotrienes, especially leukotriene D2. Representing the antagonist activity of the

of this invention, The following is a series of and the values pA ₂ and calculated at from the protocols of test above. (Π) In vitro Compounds gives Formula P 2 X mr R _o - 1 2 -C0 ₂ h 1-H -OH 6.0 -CH ₂ 0H 1-H -OH 5.3 -CHCH-OH 2 1-H -OH 5.2 OH

62 368

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I

ce

r-4

HERE

-16

ONLY κβ □ 'χ (I.e. (I.e. ισ C0 1 αο I 1 ι ω 1 00 (I.e. σ \ 1 CA σ \ 1 ΙΛ ι ι is it 1 Ch

Compounds of Formula (III) In Vitro In Vivo

O

xo

O

0)

fl

43

W

(d)

O

W

O

O

the

EC

EC |

σι σι σι σι σι σι -X JC -X and; and yc \ \ \ σι σι σι ι ι σι ι □ » e e e ι ε e ι ε (I.e. (I.e. (I.e. (I.e. (I.e. (I.e.

I σι .X σι σι .χ ι σι ι I 1 1 1 And ι ε ι 1 1 1 (I.e. or

Α 04 04 04 Ί γ-4 (I.e. α \ (I.e. .-1 Γ-1 (I.e. r-I +! νθ (I.e. HERE <± (I.e. ν (I.e. (I.e. Μ3 (I.e. (I.e. (I.e. (I.e. (I.e. (I.e. (I.e. Μ3 (I.e. (I.e. (I.e. (I.e.

X

04

O

CJ

OJ

oil

□ J

Ci

ε |

X χ X 04 X or (I.e. 04 X 2 α (I.e. (I.e. (I.e. α U ω ω ζ-X ω 04 04 04 (I.e. 04 X χ X X χ ω α u U CJ 04 X χ χ X X X X X X X X X (I.e. 2 2 2 (i.e. 2 (I.e. 2 σ or α (I.e. α 1 1 1 1 ι 1 1 1 ι ι 1 1 (I.e. 04 04 or 04 X X X X 2 2 2 2 2 X X 1 I X 1 X X 1 1 ^χ X χ X X χ X X X X X X X X X X α α α or α α α or or α α α σ ι ι ι 1 1 ¹ 4 1 I ι ¹ ι X χ χ X X χ χ X X χ χ X ί ~ ί α α or or α or or α α (I.e. or I ι ι 1 1 I ¹ 1 X (I.e. 1-1 • -Ι ι-4 ι-4 ι-4 r-4 ι-4 γ-4 - 4 hi ι-4 1-1 (I.e. (I.e. rH (I.e. ι-4 Γ-1 (I.e. ι-4 ι- | (I.e. rH rH 1-4 (I.e. (I.e. or (I.e. (I.e. 04 04 04 (I.e. (I.e. 04 ι-4 ι-4 (I.e. (I.e. ι-1 ι-4 (I.e. ι-4 α (I.e. hi rH 04 ιΗ 1-1

X

04

O

ω

or

X

CJ

XX

2 O

II

(I.e.

(I.e.

ro ro 2 2 Σ ΙΛ ro ro ·% (I.e. τι O ι-4 rH (I.e. ro ro X tn tn 2

z Z-X Z-X CD ass CT ✓

CO X)

(I.e.

ro χι

04

X

(I.e.

XXX

04 ΟΙ

ι- | ι-4 Ή

tn X i-4 ro (I.e. IA (I.e. O O z-χ (4 · » ox tn ro i-i X-tf Q. <r here

04 04 04 Μ

ι- | ι- | ι-1 γ-I

ass. σι

ο ο Ρ te ο ω Ε Ε Ο Ό 0) 0) Μ ι-I

ro χι

(c) Agonist

62 368

SKB CASE 14135-58-59

(i.e.

Α specificity of the antagonist activity of the compounds set ds dBsta relativameji invention is demonstrated by low levels of antagonism towards you as agonists Potassium sodium chloride, carbachol, histamine and ^p ^ GF _2c.

The pharmaceutical compositions of the present invention comprise a pharmaceutical carrier or diluent and an amount of a compound of formula (I) or a pharmaceutically acceptable salt, such as a sufficient alkali metal salt thereof to inhibit the effects of leukotrienes, such as the symptoms of asthma and other allergic diseases.

When the pharmaceutical composition is employed in the form of a solution or suspension, examples of suitable pharmaceutical carriers or diluents include:

for water systems, water; for non-aqueous systems, ethanol, glycerin, propylene glycol, cereal oil, cottonseed oil, peanut oil, sesame oil, liquid paraffins, and water blends thereof for the solid, lactose, kaolin and mannitol systems) and for aerosol systems, dichlorodifluoromethane, chlorotrifluoroethane and compressed carbon dioxide.

In addition to the pharmaceutical carrier or diluent, the present compositions may further include other ingredients, such as stabilizers, antioxidants, preservatives, lubricants, suspending agents, viscosity modifiers and the like, provided such additional ingredients do not have a detrimental effect on the therapeutic action of the present compositions.

The nature of the composition and the vehicle and the pharmaceutical diluent will obviously depend on the route of administration, i.e. parenteral or inhalation.

In general, especially in the case of prophylactic treatment of asthma, the compositions will be in a form suitable for administration by inhalation. Thus, the compositions will comprise a suspension or solution of the active ingredient in water for administration by a conventional vaporizer. Alternatively, the compositions will comprise a suspension or solution of the active ingredient in a conventional liquefied propellant or compressed gas for

62 368

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-18-

administration from a pressurized aerosol type container. The compositions may also comprise the solid active ingredient diluted with a solid diluent for administration from a powder inhalation device. In the above compositions, the amount of carrier or diluent will vary but will preferably be the dominant part of a suspension or solution of the active ingredient. Where the diluent is a solid, it may be present in amounts less than, equal to or greater than those of the solid active ingredient.

z

For parenteral administration the pharmaceutical composition will be in the form of sterile injectable liquid, such as an ampoule or an aqueous or non-aqueous liquid suspension.

Typically, a compound of formula I is administered to an animal in a composition comprising a nontoxic amount sufficient to cause an inhibition of the symptoms of an allergic response. When employed in this way the dosage of the composition is selected from the range of 350 mg to 70 mg of active ingredient per each administration. For convenience, equal doses will be administered 1 to 4 times a day, the daily dosage regimen being chosen from about 350 mg to about 2800 mg.

The pharmaceutical preparations thus described are made following the conventional techniques of the pharmaceutical chemist appropriate for the intended end product.

This disclosure includes the method of initiation. The present invention provides a method for the treatment of an allergic response resulting from the release of a mediator comprising administering to an animal a therapeutically effective amount for causing said inhibition of a compound of formula I, preferably in the form of a pharmaceutical composition. Administration may be done in dosage units at appropriate intervals or in single doses as needed. Usually this method will be practiced when the relief of allergic symptoms is specifically required, but can also be applied as a continuous or prophylactic treatment. It is incumbent upon those skilled in the art to determine by routine experimentation the effective dose to be administered chosen from the dose range

indicated above, taking into account factors such as the degree of pregnancy of the allergic state being treated, and so on.

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The following examples explain the preparation of the compositions of this invention and their inclusion in pharmaceutical compositions, and as such do not limit the invention set forth in the claims appended to this disclosure.

EXAMPLE 1

Preparation of 3- [2- (2-carboxyethyl) thio] -4 (Z) -heptadecen-1,2-diol

(a) Heptadec-2 (E), 4 (z) -dienyl tetrahydropyranyl ether

l (a) (l)

Heptadec-2 (E), 4 (E) -dienyl ether tetrahydropyranyl ether

Κ ·) (2)

Tridecyltriphenylphosphonium bromide (189 g, 0.3 mol) was dissolved in 900 ml of tetrahydrofuran and cooled to 0 ° C in an ice-salt bath with stirring under argon. A 2,2 N solution of n-butylthio in hexane (250 ml, 0.36 mol) was added dropwise over 30 min. The mixture was stirred for another 20 min. and then cooled to -70 ° in a dry ice-bath, -acetone. 4-Hydroxybut-2 (E) -en-1-al tetrahydropyranyl ether (51 g, 0.3 mol) in 225 ml of tetrahydrofuran was added dropwise over 35 min. and the mixture was stirred for an additional 1 hour at -70 DEG. The mixture was then poured into 6.25 liters of ether and stirred for 20 minutes. The resulting mixture was filtered through glass fiber filter paper. The filtrate was evaporated and the residue was triturated with hexane, filtered and evaporated to give a 3: 1 mixture of 1 (a) (1): 1 (a) (2).

(b) Heptadec-2 (E), 4 (Z) -dien-1-ol 1 (b)

Heptadec-2 (E), 4 (E) -dien-1-ol 1 (b) (2)

The mixture of compounds 1 (a) (1), (a) (2)

(80 g, 0.24 mol) in 3 liters of methanol was added pyridinium p-toluenesulfonic acid (3 g, 0.012 mole) to the stirring mixture under argon at room temperature. The reaction was monitored by TLC. After completion of the reaction, the solvent was evaporated and the residue was flash chromatographed on 500 g of silica gel, washing with 10% ethyl acetate in hexane to give a 3: 1 mixture of 1 (b) ( l): 1 (b) (2). Separates

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-20-

(b) (l) of l (b) (2) was carefully chromatographed on silica gel. Melting point of compound 1 (b) (1): 342-370. Melting point of compound 1 (b) (2): 512-552.

(c) Trans-2,3-epoxyheptadec-4-Z-ene-1-ol (c)

Compound 1 (b) (1) (2.52 g, 10 mmol) was dissolved in 100 mL of methylene chloride stirring at room temperature in vac. bead of argon. A 0.5N solution of sodium bicarbonate (30 ml) was added. The acid was slowly added in small portions. of 85% m-chloroperbenzoic acid (2.03 g, 10 mmol). The mixture was stirred for 1.5 hours after the addition was complete. The phases were separated and the aqueous layer was washed with methylene chloride. The combined organic phases were dried over anhydrous sodium sulfate, filtered and evaporated. The residue was chromatographed instantly on silica (100 g), washing with 10-20% ethyl acetate / hexane to provide compound 1 (c).

(d) 3 - [(2-carbomethoxyethyl) thio] -4 (Z) -heptadecen-1,2-diol (d)

Compound 1 (c) (7.2 g, 26.9 mmol) was dissolved in 40.2 ml of methanol containing 2% of triethylamine. This solution was stirred at ambient temperature under argon and a solution of methyl 3-mercaptopropionate (4.92 mL, 44.4 mmol) and triethylamine (11.16 mL, 80.2 mmol) in 40 , 2 ml of methanol dropwise over 15 minutes. The mixture was stirred for 5 hours at room temperature and then placed in the refrigerator, leaving it overnight. The solvents were evaporated and the residue was flash chromatographed on 500 g of silica gel washing with 10-50% ethyl acetate in hexane to give 1 (d), mp 33-360,

(±) -3- (2-carboxyethyl) thio] -4 (Z) -heptadecen-1,2-diol 1 (e)

Compound 1 (d) (500 mg, 1.29 mmol) was dissolved in 2 mL of methanol and stirred under argon. An IN solution of sodium hydroxide (0.65 mL, 0.65 mmol) was added dropwise and the mixture was stirred for 0.5 h at room temperature. An additional 0.65 ml of 1N NaOH solution was added and the mixture was stirred for 1 hour. TLC indicated that there was still some starting material, thereby adding more 0.2 g

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of 1N NaOH solution and the mixture was stirred for an additional 1 hour and half. The methanol was stripped and the residue acidified with dilute hydrochloric acid. The mixture was extracted with diethyl ether and the organic phase was dried over anhydrous magnesium sulfate, filtered, and evaporated to give crude product. This was recrystallized from diethyl ether / hexane to give the desired compound, m.p. 76-77Â °.

Boric acid Found W 64.13 64.39 H 10.23 10.22 s 8.56 8,78

EXAMPLE 2

Preparation of 2- [2- (2-carboxyethyl) thio] -3 (Z) -hexadecen-1

(a) 2 - [(2-carbomethoxyethyl) thio] -3 (Z) -hexadecen-1-al2 (a)

Compound 1 (d) (2 g, 5.15 mmol) was dissolved in 10 mL of diethyl ether and stirred in a water bath at room temperature. Saturated solution (100 ml) of periodic acid in diethyl ether was added in one portion. The resulting mixture was stirred for 2 min. and then immediately flash chromatographed on 150 g of silica gel with 10% ethyl acetate in hexane affording 2 (a).

(b) 2- [2- (2-carbomethoxyethyl) thio] -3-3 (z) -hexadecen-1-ol

The compound of Example 2 (a) (192 mg, 0.5 mmol) was dissolved in 1 ml of methanol and stirred at 0 ° under argon. Sodium borohydride (38 mg, 1 mmol) was added and the mixture was stirred for 5 minutes. The mixture was acidified with dilute hydrochloric acid and evaporated to dryness. The residue was taken up in diethyl ether and washed with dilute hydrochloric acid, aqueous sodium bicarbonate, dried over anhydrous sodium sulfate, filtered, and evaporated to give crude product which was flash chromatographed was dissolved in 20 g of silica gel washing with 15% ethyl acetate in hexane to give the above compound.

(c) 2- [2- (2-carboxystyl) thio] -3 (Z) -hexadecen-1-ol

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The compound of Example 2 (b) (180 mg, 0.5 mmol) was dissolved in 1 mL of methanol and 1N sodium hydroxide solution (1 mmol, 1 mmol) was added. The mixture was stirred at ambient temperature under argon for 3 hours and then placed in the freezer over the weekend and then stirred for a further 2 hours at room temperature until tlc indicated that there was only one trace of the starting material. The methanol was stripped and the residue acidified with dilute hydrochloric acid, and extracted with chloroform. The organic phase was dried over anhydrous magnesium sulfate, filtered, and evaporated to give crude product. This was recrystallized from hexane at -78Â ° C and the desired compound, mp 39-40Â °, was obtained.

Theoretical Found W 66.23 66.47 H 10.53 10.77 s 9.30 9.15

EXAMPLE 3

Preparation of 2 - [(2-carboxyethyl) thio] -3 (Z) -hexadecarboxylic acid

cenic

(a) 2- (2-Carbomethoxyethyl) thio] -3 (Z) -hexadecenoic acid 3 (a)

Methyl 2- (2-carbomethoxyethyl) thio] -3 (Z) -hexadecenoate

3 (.) (2)

The compound of Example 2 (a) (0.8 g, 2.25 mmol) was dissolved in 5.5 mL of acetone. The solution was cooled to -40Â ° C and stirred under argon. Oones reagent (0.3 ml, 0.6 mmol) was added and the mixture was stirred at -302 g - 40 ° C for 30 minutes. Additional Oones reagent (0.3 ml, 0.6 mmol) was added and the mixture was stirred for 1 hour. The remaining Oones reagent (0.145 mL, 0.29 mmol) was added. Half an hour later, the reaction was stopped and maintained at a temperature range of -300 ° -40 ° C by adding 1 ml of isopropanol. He took up the solvents and the residue partitioned in diethyl ether and H ₂ 0. extracted the aqueous phase with diethyl ether. The combined organic phases were dried over anhydrous MgSO4, filtered and evaporated to give crude product 3 (a) (1). In order to facilitate

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After purification, compound 3 (a) (1) was treated with an excess of diazomethane in diethyl ether to DS and allowed to warm to room temperature. The solvents were evaporated and the residue was flash chromatographed on 200 g of silica gel washing with 5% EtOAc in hexane to give the desired product 3 (a) (2).

(b) 2- [2- (2-Carboxyethyl) thio] -3 (Z) -hexadecenoic acid

Compound 3 (a) (2) (0.16 g, 0.41 mmol) was dissolved in

3.2 ml of methanol, and stirred under argon at 0 ° C. Adi. A 1N sodium hydroxide solution (1.6 mL, 1.6 mmol) was added and the ice bath was removed. The mixture was stirred at room temperature for 2.5 hours. The methanol was evaporated and the residue was cooled in an ice bath and acidified with dilute HCl. The aqueous phase was extracted twice with diethyl ether. The combined ether extracts were dried over anhydrous sodium sulfate, filtered, and evaporated to give crude product. This was recrystallized from diethylhexane / ether and the desired compound was obtained, mp 52-54 ° C. Calculated composition: C 63.65; H 9.56; S, 8.94. Found: C 63.59; H 9.45; S, 9.24. EXAMPLE 4

Preparation of 5- / ~ (2-amino-3-carboxymethylamino-3-oxopropyl) thio J7-4 _i-hydroxy-6 (Z) -nonadecenâico. such as 4 (R), 5 (S) isomer, 4 (S), 5 (R) isomer and the mixture of the two isomers.

(a) 3-Carbomethoxypropionyl chloride 4 (a)

To a glacial solution of monomethyl succinate (150 g, 1.135 mol) in 900 ml of methylene chloride and 3 ml of Î ±, β-dimethyl formamide was added 119 ml (1.26 mol) of oxalyl chloride, maintaining the pot temperature below 52 ° C. After the addition was complete, the reaction mixture was stirred at 0 DEG C. for 1 hour and then concentrated in vacuo to give a crude yellow liquid which was used without purification in the Rosenmund reduction.

(b) 3-Carborethoxypropionaldehyde 4 (b)

To the acid chloride 4 (a) (8.9 g, 0.059 mol) in 200 ml of sieve dried tetrahydrofuran was added 6.9 ml (0.059 mol)

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-24-

of 2,6-lutidine. After a half hour of standing at room temperature, the mixture was filtered and 0.7 g of 10% palladium on carbon was added to the filtrate. The mixture was hydrogenated at 50 psi for 2 hours, the solids were removed by filtration and the filtrate was concentrated. The residue was dissolved in methylene chloride, washed twice with 10% hydrochloric acid solution and then twice with 5% sodium bicarbonate solution. The organic extract was dried over anhydrous sodium sulfate and concentrated in vacuo. Distillation in bead tube (40-558 ° C / 0.05 mm) gave a colorless liquid.

(c) 5-carbomethoxy-2-pentenal 4 (c)

To a mechanically stirred solution of 4 (b) (72.1 g, 0.620 mol) in 750 mL of toluene under argon was added formylmethylene triphenylphosphorane (235.9 g, 0.776 mol). The reaction mixture was refluxed for 1.5 hours, cooled to room temperature and then concentrated in vacuo. Dei. The residue was dissolved in diethyl ether at 0 ° C overnight. The mixture was filtered and the solid was washed with cold water. The filtrate was concentrated and the resulting brown oil was subjected to distillation on a bead tube. The product was collected at 65-80 ° C / 0.05 mm,

(d) methyl 4,5-epoxy-6-oxohexanoate 4 (d)

To a solution of 36.4 ml of a 30% solution of hydrogen peroxide and 58.2 ml of 1N sodium bicarbonate in 800 ml of methanol and 400 ml of water under argon was added dropwise was added 41.2 g (0.29 mol) of 4 (c) in 400 ml of methanol over 45 minutes. After the addition was complete, the reaction mixture was stirred for 2.5 hours at room temperature, maintaining the pH between 9 and 9.5 by the addition of sodium bicarbonate solution. The reaction mixture was then poured into 1 liter of saturated ammonium sulfate solution, the methanol removed in vacuo, the solids removed by filtration and the product extracted into methylene chloride. The aqueous layer was subsequently extracted twice, the combined extracts were dried over anhydrous sodium sulfate and then concentrated to give a crude golden oil. The product was distilled in a bead tube at 82-98 ° C / 0.1 mm.

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-25 (θ) 4,5-epoxy-6 (Z) -nonadecenoate 4 (e)

To a glacial solution of tridecyltriphenylphosphonium bromide (preparation described below) (97.2 g, 0.185 mol) in 600 ml of tetrahydrofuran dried in an argon atmosphere was added dropwise 17.1 ml of a solution 2 , 4M n-butyl. uncle in hexane ; the temperature in the ^Q-manteue C during the addition of 30 minutes. The reaction mixture was stirred at this temperature for a further 15 minutes, cooled to -78Â ° C (dry gell / isopropanol) and then 26.5 g (0.168 mol) of 4- ( d) in 150 ml of dried tetrahydrofuran. After the addition was complete, the reaction was stirred for 1 hour at -78 ° C and then concentrated in vacuo at 24 ° C. The residue was triturated with hexane and then allowed to stand at 0Â ° C overnight. The hexane was decanted and then the residue was sonicated four times with hexane. The combined extracts were concentrated in vacuo and the crude product was purified by preparative HPLC (2 columns of Waters Prepak silica, washing with 8% ethyl acetate in hexane) to give 4 (e) in the form gives an oil.

(f) Tridecyltriphenylphosphonium bromide

A solution of 1-bromotridecane (100 g, 0.4 mol) and triphenylphosphine (100 g, 0.4 mol) in 500 ml of xylenes was refluxed overnight. The reaction was then cooled to room temperature and poured into diethyl ether. The resulting oil was washed three times with ether, dissolved in methylene chloride and then concentrated in vacuo. Dei. The oil was then allowed to stand in diethyl ether at 0 DEG C. overnight and then concentrated in vacuo to give a white solid.

(2-trifluoroacetamido-3-carbomethoxymethylamino-3-oxopropyl) thio] -6 (Z) -nonadecenoic acid methyl ester

To epoxide 4 (e) (12.9 g, 0.04 mol) was added dropwise

dropwise over 1 hour under an argon atmosphere and at room temperature.

a solution of 14.5 g (0.55 mol) of 2-trifluoroacetamido-362

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0

-carbomethoxymethylamino-3-oxo-propylmercaptan.

(HSCH ₂ CH ₂ C0 ₂ ÇHtNHCH _j)

in triethylamine (6.7 ml) / methanol (150 ml). The reaction mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was dissolved in a minimum volume of methylene chloride, 100 ml of hexane was added and this solution was then stored at -158C for 1 hour. The resulting solid was filtered off and then the filtrate was concentrated in vacuo to give a crude oil which consisted of a mixture of the desired products.

(h) | "acid lactone 4 (R) -hydroxy-5 (S) - _i /""(2-trifluoroacetamido-3-carbometoximetilamino-3-oxopropyl) tio_7 -6 (Z) -nonadecenoic acid and the lactone 4 (S) -hydroxy-5 (R) - [(2-trifluoroacetamido-3-carbomethoxymethylamino-3-oxopropyl) thio] -6 (Z) -nonadecenoic acid

The crude mixture of Example 4 (g) (20.4 g)

g) in 200 ml of toluene in the presence of 75 mg of p-toluenesulfonic acid for 15 minutes. The reaction mixture was then concentrated in vacuo and the resulting crude oil was purified by preparative HPLC (2 Waters Prepak silica columns, washing with 45% ethyl acetate in hexane) to give the desired products as the isomer 4 (R), 5 (S) pure and the pure 4 (S), 5 (R) isomer.

(i) hydrate of 4- (R) -hydroxy-5 (S) - / "" (2-amino-3-carboxymethyl - amino-3-oxopropyl) tio_7-6 (Z) -nonadecenoic acid ^{50 *}

The lactone of Example 4 (h) as the 4 (R), 5 (S) -isomer (1.9 g, 0.00327 mol) in aqueous sodium hydroxide ( 0.72 g, 0.018 mol in

50 ml of water). The pH of the reaction mixture was then adjusted to

3.5 with concentrated hydrochloric acid and collected and dried the resulting solid: mp 144-146Â ° C. Calculated for

^C 24 ^H 44 ^N 2 ^O 6 ^S * ^{1 l and / 4 H} 2 ^{O C 56, 38, H 9, 17} ? ^{5 C} '· ⁴⁸ Found:

C 56.16; H 8.83; N, 5.54.

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The lactone of Example 1 (h) was further converted as the 4 (S), 5 (R) isomer into the desired 4 (S), 5 (R) -isomer of the above compound mp 141-1439 ° C; comp. calculated for .1 1/4 H ₂ 0: C , 56.39; H 9.17; N 5.48. Found: C 56.20;

H 8.88; N, 5.18.

The isomeric mixture of the desired product was obtained by treating the product mixture prepared according to Example 1 (g) without further purification as described above: mp 140-1432 ° C. Comp. calcd for 4 ^ 4 ^ ^^ ^{χ 5.1} / ^{4 56 C Η θί} 2 ^{'39I H 9' 5 N 17j} 'of ⁴⁸ · are: C 56.44; H 8.75; N, 5.16.

EXAMPLE 5

Preparation of 4-hydroxy-5 - [(2-carboxyethyl) thio] -6 (Z) -nonadecenoic acid as a mixture of the 4 (R), 5 (S) and 4 (3), 5 (R) .

A solution of 1.32 g (4.1 mmol) of 4,5-epoxy-6 (Z) -nonadecenoic acid methyl ester and 1.46 g (12.2 mmol) of 3-mercaptopropionate methyl ester in 20 ml of methanol was treated with 1.62 g (16.1 mmol) of triethylamine in 10 ml of methanol under argon overnight. The reaction mixture was concentrated in vacuo and purified by silica gel chromatography with hexane / ether (60/40) to give the diester containing a little of the sulfonate / lactone as evidenced by IR and TLC ( CH ₂ Cl ₂ / hexane / acetone, 47/47/5).

A solution of this diester / lactone mixture (1.6 g, 3.6 mmol) in 30 mL of methanol was treated with 1.43 g (36 mmol) of sodium hydroxide in 5 mL of water at ambient temperature and under an argon atmosphere. The reaction mixture was partially concentrated in vacuo, redissolved in 20 mL of water and acidified with dilute phosphoric acid. The aqueous solution was extracted three times with 50 ml of ether and the extracts were dried over magnesium sulfate, filtered and concentrated in vacuo to give a white crystalline solid, mp 78 DEG-80 DEG C., calculated for Calc'd for C 20 H 13 NO 4: C 63.43; H 9.68. Found: C 63.11; H, 9.69.

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EXAMPLE 6

Preparation of 5 - [(3-carboxymethylamino-3-oxopropyl) thio] -4-hydroxy-6 (Z) -nonadecanoic acid.

This compound was prepared by the general method of Example 5 using 3-carboxymethylamino-3-oxopropyl mercaptan prepared by the 3,3'-dithio-dipropionic acid linkage reaction with DCC, followed by DCC The reduction of bisulfide with tri-n-octylphosphine in a 1: 1 mixture of acetone / water. The product was obtained as an oil. calc'd for C 24 H 43 N 4 O 6 ^: C 60.85; H 9.15; N 2.95. Found: C 60.65; H 9.07; N, 2.89.

EXAMPLE 7

Preparation of 5 - [(3-carboxymethyl-N-methylamino-3-oxopropyl) thio] -4-hydroxy-6 (Z) -nonadecenoic acid.

This compound was prepared by the general method of Example 5 using 3-carboxymethyl-N-methylamino-3-oxopropyl mercaptan. Racercaptan was prepared by the reaction between p-methoxybenzylthiopropionic acid and N-methyl glycine methyl ester in the presence of DCC, followed by treatment with HF at -78 ° C. 0

The product obtained had a melting point of 80-82Â ° C. Comp. calculated: C 61.57; H 9.30; N, 2.87. Found: C 61.55; H 9.65 N 3.02.

EXAMPLE 8

Preparation of 5 - [(carboxymethyl) thio] -4-hydroxy-6 (Z) -nonadeceneic acid.

This compound was prepared by the general method of Example 5, employing commercial methyl thioglycolate: mp 50-52Â ° C. Comp. calculated for ^C 21 ^H 38 ^{si C} ^ 5 & ² '; H ⁶ ^ 9.51. Found:

C 62.74; H 9.57.

EXAMPLE 9

Preparation of 5- [3- (3-carboxypropyl) thio] -4-hydroxy-6 (Z) -nonadeceneic acid.

This compound was prepared by the general method of Example 5 using commercial 4-mercapto-butyric acid, mp 56-58 ° C.

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-29Comp. calc'd for C 23 H 42 N 4 O 2 ^: 65.48; H 9.96. Found: C 63.29, H 9.75.

EXAMPLE 1Q

Preparation of 5 ^{- [} [2- (aminocarbonyl) ethyl] thio] -4-hydroxy-6 (Z) -nonadecenoic acid.

This compound was prepared by the general method of Example 5 using 2- (aminocarbonyl) ethyl mercaptan. The mercaptan was prepared by treatment of dithiocarboxylic iodide with concentrated ammonium hydroxide followed by reduction of the disulfide with tri-n-octylphosphine and 1: 1 mixture of acetone / water. The product obtained had a mp of 84-86Â ° C. Comp. calculated for ^C 22 ^H 41 ^N0 4 ^{Si 63 C 'H 9 57j' 94} · Found: C 63.56; H 9.82. EXAMPLE 11

Preparaç3o 4-hydroxy-5- / ^_ acid (2-amino-2-carboxyethyl) tio_7-6 (Z) -nonadecenoic acid as the 4 - isomer (R), 5 (S) isomer and the 4 (S), 5 ( R).

(a) 4-Hydroxy-5 - [(2-trifluoroacetamido-2-carbomethoxyethyl) thio] -6 (Z) -nonadecenoic acid methyl ester and β-

To the epoxide, methyl 4,5-epoxy-6 (z) -nonadecenoate (4e) (0.9 g, 0.00277 mol) under argon and at room temperature was added dropwise a solution of 1- , 2 g (0.00519 mol) of 2-trifluoroacetamido-2-carbomethoxyethylmercaptan in triethylamine (0.88 ml) / methanol (15 ml). The reaction mixture was stirred at room temperature for 30 hours, concentrated in vacuo and then filtered through a pad of silica gel, elutriating the product mixture with chloroform. The chloroform wash was concentrated to give a mixture of the above methyl ester and the above Y "-lactone.

(b) 4-Hydroxy-5 - [(2-trifluoroacetamido-2-carbomethoxyethyl) t- (Z) -nonadecenoic acid p-

The crude mixture (2 g) of methyl ester and α-lactone

of Example 11 (a) in 75 ml of toluene was heated at 80 ° C in the presence of 27 mg of p-toluenesulfonic acid for 20 minutes.

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The reaction mixture was then concentrated in vacuo and the residue was taken up in methylene chloride, washed with 5% sodium bicarbonate solution, dried with anhydrous brine and sodium sulfate and then concentrated in vacuo. vacuum. The resulting crude oil was applied to the preparative Water CLAP (2 columns of Prepak silica, washing with 25% ethyl acetate in hexane) to give the desired products as the 4 (R), 5 (S) - isomer and isomer 4 (S), 5 (R). Further purification of each diastereomer was carried out in a colu. on silica flash, washing with 30% ethyl acetate in hexane.

(c) 4 (R) -hydroxy-5 (S) - [(2-amino-2-carboxyethyl) thio] -6 (Z) -nonadecenoic acid sodium salt

A partial suspension of the 4 (R), 5 (S) -isomer of the β-lactone of Example 11

(b) (0.3 g, 0.00057 mol) was aqueous sodium hydroxide (0.205 g, 0.004 mol in 13 ml water). The pH of the reaction mixture was then adjusted to 3.5 with concentrated hydrochloric acid to give the desired product, mp 168-170 ° C. Comp. calculated for C ₂₂ H _{4 R, 5} S Na N 0 (H) C 58.12 H 8.87}} N 3.08. Found: C 57.99, H 8.85, N 3.70.

(d) 4 (S) -Hydroxy-5 (R) - [(2-amino-2-carboxyethyl) thio] -6- (z) -nonadecenoic acid

The reaction was carried out as described in Example 11 (c) to give the desired product, mp 159-161 ° C. Comp. calculated for

^C 22 ^H 41 ^N0 5 ^{S 1.5} * ^Na (- ^{1 '56 C 5' 69I H 8 '54' 'N3> 01} · Found: C 56.40 H 8.19}} N 3.33.

EXAMPLE 12

Preparation of 5 '' - (2-amino-3-carboxamidomethylamino-3-oxopropyl) thio] -4-hydroxy-6 (Z) -nonadecenoic acid

(a) 4 (R) -hydroxy-5 (S) - [(2-trifluoroacetamido-3-carbamoylmethylamino-3-oxopropyl) thio] -6 (z) -nonadecenoic acid, γ-lactone-4 (R) -hydroxy -5 (S) - [(2-trifluoroacetamido-3-carbamoylmethylamino-3-oxopropyl) thio] -6 (Z) -nonadecenamide

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A mixture of the lactone / methyl ester prepared according to Example 4 (g) (0.41 g, 0.7 mmol) in 10 ml of dimethoxyethane, 10 ml of methanol and 20 ml of concentrated ammonium hydroxide solution. The mixture was neutralized in the cold with concentrated hydrochloric acid to pH 7.5. The crude product was partitioned between methylene chloride and methylene chloride. The aqueous phase was further extracted twice with 40 ml of methylene chloride each time. The combined extracts were washed with brine, dried over sodium sulfate and evaporated to give an oily residue. Flash chromatography (silica gel, 1.5 ^M x 6 ^M , 3 / CH 2 OH / CHCl 3, 25 mL fraction) provided the desired products. A 6 (z) -nonadecenamide thi. has the following properties: mp 143-145 ° C; comp. calc'd for C 26 H 45 N 3 O 4 • 5 • 5 • 5, H 7.78; N, 9.61. Found: C

53.86; H 7.52; N 9.83.

(b) 4 (R) -hydroxy-5 (S) - / ^_ (2-amino-3-carbamoylmethylamino-3-oxopropyl) tio_JZ-6 (Z) -nonadecenoic acid

A mixture of 120 ml (0.2 mmol) of the V-lactone of Example 12 (a) in 5 ml of 0.2 M sodium hydroxide solution was stirred at ambient temperature for 18 hours. You acidified yourselves. to pH 3 by adding a concentrated solution of hydrochloric acid in an ice bath. The resulting precipitates were filtered and washed rapidly with cold water and dried at 56Â ° C for 24 hours to give the desired product, mp 143-145Â ° C. Comp. calculated: C 57.51; H 9.35; N, 8.38. Found: C 57.50; H 9.42; N, 6.72.

EXAMPLE 13

Preparation of 3-hydroxy-4 - [(2-carboxyethyl) thio] -5 (Z) -octadecenoic acid

(a) methyl 3- (1,2-dihydroxyheptadec-4 (Z) -enyl) thio-propionate

13 (a)

Compound 1 (c) (7.2 g, 26.9 mmol) was dissolved in 40.2 g

ml methanol containing 2% triethylamine. This solution was stirred at ambient temperature under argon, and

was added dropwise over 15 minutes a solution of 3-mercaptoprop-368

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(4.92 mL, 44.4 mmol) and triethylamine (11.16 mL,

80.2 mmol) in 40.2 ml of methanol. The mixture was stirred for 5 hours at room temperature, and then refrigerated overnight. The solvents were evaporated and the residue was flash chromatographed on 500 g of silica gel, washing with 10-15% ethyl acetate in hexane to give 13 (a), mp 33-36 ° C,

(b) 2- [2- (carbomethoxyethyl) thio] hexadec-3 (z) -enal 13 (b)

Compound 13 (a) (2 g, 5.15 mmol) was dissolved in 10 mL of diethyl ether and stirred in a water bath at room temperature. A saturated solution (100 ml) of periodic acid in diethyl ether was added one time. The resulting mixture was stirred for 2 minutes and then flash chromatographed on 150 g of silica gel with 10% ethyl acetate in hexane to give 13 (b).

(o) Methyl 3-hydroxy-4- [2- (2-carbomethoxyethyl) thio] octadec-5 (Z)

A dry sealed flask was charged with 4.5 ml of hexane and maintained under an argon atmosphere, and cooled in an ice bath. A 2.2M solution of n-butyl lithium (1.03 mL, 2.25 mmol) was added dropwise followed by dropwise addition of diisopropylamine (0.315 mL, 2.25 mmol). The solution was stirred at 80 ° C for 10 min. and then cooled to -78 ° C in a ge. dry acetone for 15 minutes. It was added over 1 min. to a solution of methyl acetate (0.18 mL, 2.25 mmol) in 1.5 mL of hexane and the mixture was stirred at -78 ° C for an additional 1 minute. At this point the mixture was almost transparent. Compound 13 (b) (750 mg, 2.1 mmol) was added over 1 minute in

1.5 ml of hexane, which gave a clear yellow solution which was stirred at -78 ° C for an additional 15 minutes. The reaction mixture was then flash chromatographed on 100 g of silica gel washing with 15% ethyl acetate in hexane, giving compound 13 (c).

(d) 3-Hydroxy-4 - [(2-carboxyethyl) thio] -5 (Z) -octadecenoic acid

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Compound 13 (c) (0.2 g, 0.46 mmol) was dissolved in 5 mL of methanol and stirred under argon at &lt; 0 &gt; C. A 1 N sodium hydroxide solution (2 ml, 2 mmol) was added dropwise over half a minute. The ice bath was removed and the reaction was allowed to warm to room temperature for 2 hours. Most of the methanol was evaporated and the aqueous residue cooled in an ice bath, acidifying with dilute hydrochloric acid. The aqueous phase was extracted twice with diethyl ether. it. The combined ether extracts were dried over anhydrous sodium sulfate, filtered, and evaporated to give crude product. This was recrystallized from diethyl ether-hexane and the desired compound was obtained, mp 88-97Â ° C. Comp. Calc'd: C 62.65; H 9.51% S 7.96. Found: C 62.32; H 9.38; S, 8.10. EXAMPLE 14

Preparation of 5- (2-carboxyethyl) thio-7- (Z) -nonadecenoic acid

(a) 2-Bromo-5-carbomethoxypentanoic acid 14 (a)

90 g of thionyl chloride was added to a solution of 30 g (0.187 M) of 5-carbomethoxypentanoic acid in 250 ml of chloroform. The mixture was refluxed for 2 hours, cooled and the solvent was removed in vacuo. The residue was redissolved in 200 ml of carbon tetrachloride, 40 g (0.225 M) of N-bromosuccinimide was added, the mixture was heated to reflux, eight drops of a mixture of HBr / acetic acid, and the reaction was refluxed for 3 hours. After cooling, the mixture was filtered and evaporated to dryness. The residue was dissolved in 250 ml of acetone, to which was added dropwise 200 ml of a 5% solution of sodium bicarbonate prepared just before. After 15 min. the period in which the pH was constantly adjusted to 9 with sodium bicarbonate, the mixture was acidified with dilute HCl to a pH of 1.0, concentrated in vacuo and extracted with chloroform. The organic phase was extracted with dilute bicarbonate solution, acidified and extracted with chloroform. The extract was dried over MgSO4, filtered and evaporated to dryness to give the desired compound as a viscous oil.

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(B) 5-Carbomethoxy-2 - [(methoxycarbonylpropyl) thio] pentanoic acid 14 (b)

A mixture of 42 g (0.175 M) of 14 (a), 50.68 g (0.42 M) of methyl 3-mercaptopropionate and 147 ral (1.05 M) of triethylamine was refluxed for two hours in 250 mL methanol, cooled, and concentrated in vacuo. The residue was dissolved in ethyl acetate and extracted with 5% sodium bicarbonate. The aqueous extract was acidified with 3N HCl, extracted with dichloromethane, dried over MgSO 4, filtered and evaporated to dryness to give the desired compound.

(c) 5-carbothoxy-2 '- (methoxycarbonylpropyl) thio] pentanol 14 (c)

A solution of 14 (b) (40 g, 0.142 M) in THF (200 ml) was placed in a methanol-ice bath (-10 DEG C.). The contents were allowed to cool for 20 min, at which time it was slowly added for 50 min. 12.57 g (0.156M) borane-methyl sulfide complex, then refluxing for 30 minutes. Acetic acid (10 mL) was then added and excess THF was evaporated. The reaction mixture was dissolved in ethyl acetate (300 ml), washed with 5% sodium bicarbonate, dried over MgSO 4 and filtered. Evaporation gave the crude alcohol.

(d) 5-carbomethoxy-2 '- (methoxycarbonylpropyl) thio] -pentanal 14 (d)

Were combined and cooled to -652C act methylene chloride (10 ml) and S0 ₂ Me (l 78 g, 0.23 M). TFAA (1.73 g, 0.015 M) was added dropwise to the cold solution, at which point a precipitate formed. After 5 minutes at -65 ° C a solution of 14 (c) (2.0 g, 7.58 mmol) in CH ₂ Cl ₂ (5 mL) was added dropwise, maintaining the reaction mixture at - 652C. The mixture was then stirred at -65 ° C for 30 minutes, then TEA (2.30 g,

0.023 M) dropwise. A temperature below -60Â ° C was maintained until the addition of TEA was complete. The bath was then removed and the reaction was allowed to warm to room temperature. The mixture was diluted with CH ₂ Cl _2, washed with 3N HCl, H ₂ O twice with 5% NaHCO 3 and once with brine. The organic phase was then dried over MgSO 4, filtered and evaporated to dryness to give the crude aldehyde.

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(e) methyl 5 - [(2-carbomethoxyethyl) thio] -6 (Z) -nonadecanoate

Mai

A mixture of tridecyltriphenylphosphonium bromide (1.76 g, 3.35 M) and THF (20 ml) was stirred for 5 minutes at room temperature. The solution was then cooled to -6 ° C and stirred for an additional 10 minutes at which time n-butylthio (3.05 mmol) was added slowly maintaining a temperature of -65 ° C. After stirring for 10 min at -89 ° C, the reaction mixture was stirred for an additional 10 minutes at -1 ° C. After recooling to -89 ° C 14 (d) (0.918 g, 3.50 mmol) in THF (10 mL) was dissolved and slowly added. The reaction was stirred for 1 hour at -69 ° C, then the bath was removed to warm the mixture to room temperature. The THF was then evaporated and the residue was dissolved in ethyl acetate, washed with 3N HCl, water and twice 5% NaHCO 3. The extract was dried over MgSO4, filtered and evaporated to dryness. The product was chromatographed sequentially with 91% hexane: 9% EtOAc to give the nonadecanoate.

(f) 5- [2- (2-Carboxyethyl) thio] -6 (Z) -nonadecanoic acid

Potassium carbonate (0.81 g, 5.84 mmol) was dissolved in H ₂ O (10 mL) to which was added methanol (5 mL). The solution was stirred for 2 minutes at room temperature, then 14 (s) (100 mg, 0.234 mmol) in MeOH (24 mL) was added. The mixture was allowed to stir overnight at room temperature, whereupon the excess MeOH was evaporated and the remaining aqueous phase was washed with ethyl acetate, acidified to a pH of 1.0 with 3N HCl and extracted three times with ethyl acetate, the extract dried over MgSO 4, filtered and evaporated to an oil. The product was recrystallized from petroleum ether-ether and cooled for 1 hour to give the nonadecanoic acid as a white crystalline solid, mp 53-54 ° C.

EXAMPLE 15

Preparation of 5 ^{- [} (2-carboxyethyl) thio] -4-hydroxy-6 (Z) -ureanamide.

62 or 36

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-36-

This compound was prepared by treating the compound of Example 5 with 1% trifluoroacetic anhydride in CH ₂ Cl ₂ for 2 hours at room temperature, followed by treatment with NH ₃ in methanol solution at 0 ° C for half an hour and at the temperature from one day to the next. The product was obtained as a crystalline solid, mp 82-85 ° C. Comp. Calc'd for C 20 H 32 N 3 O 4 • NH 4 • 0.3M: C 62.77; K 9.89; N, 4.33. Found: C 62.67, 62.98; H 9.92 9.78; N 3.98 4.00.

EXAMPLE 16

Preparation of 5- (3-carboxymethylamino-3-oxopropyl) thio] -4-hydroxy-6 (Z) -nonadecenamide

A mixture of 0.9 g (1.4 mmol) of the compounds of Example 4 (g) in 100 ml of 3.8% anhydrous ammonia in ethanol solution was stirred at ambient temperature for two days. The reaction mixture was concentrated to dryness. The residue was azeotroped with methylene chloride to give 0.9 g of off-white powder. The hydroscopic material was dissolved in 10% NaOH solution (5 mL) and chromatographed on a 3 cm x 9 cm column of XAD-7 resin. After washing with H ₂ O (100 mL) the column was washed with aqueous methanol (1: 1) dropping 20 mL per fraction. Fraction 13 was collected and the desired product obtained as a white amorphous powder, mp 195-19 ° C. Comp. Calc'd for ^C 24 ^H 44 ^N 3 ^O 5 ^S * Na (H) 2 ₂ O: C 52.82; H 8.87; N 7.70. Found: C 52.65;

H, 8.15; N, 7.72.

EXAMPLE 17

Preparation of 4-hydroxy-5 - [(2-carboxyethyl) thio] - (Z) -hepadenoic acid

This compound was prepared by the general method of Example 5 using methyl ester of 4,5-epoxy-6 (Z) co -heptadecenói _1, obtained by employing the general method of Example 4 (e) with undeciltrifenilfosfónio bromide. The product obtained had a melting point of 78.5-80Â ° C. Comp. calculated: C 61.82; H 9.34; S, 8.25. Found: C 61.82; H 9.10; S 8.42.

EXAMPLE 18

Preparation of 4-hydroxy-5 - [(2-carboxyethyl) thio] -6 (Z) -hexenoic acid

62 368

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-37- ΠΜρΗ

This compound was prepared by the general method of Example 5 employing 4,5-epoxy-6 (z) -heenoicosenoic acid methyl ester, obtained by the general method of Example 4 (e) employing pentadecyltriphenylphosphonium bromide. The product obtained had a melting point of 59-61 ° C. Comp. Calc'd: C 64.82; H 9.97; S, 7.21. Found: C 65.16; H 10.07; S, 7.36.

EXAMPLE 19

Preparation of 4- [2- (2-carboxyethyl) thio] -5 (Z) -octadecenoic acid.

(a) methyl 4-bromo-4-carboxybutanoate 19 (a)

Methyl 4- (chloroformyl) butyrate (30.0 g, 0.182 M) was introduced into a flask along with 200 ml CCl3. To this solution was added N-bromosuccinimide (40.0 g, 0.219 M). The reaction mixture was refluxed and seven drops of 47% aqueous HBr were added, the conditions of reflux continued for 3 hours. of 2 hours. After cooling to room temperature, the reaction was filtered and evaporated to dryness. The residue was then left in the refrigerator overnight. The residue was dissolved in 250 ml of acetone, saturated sodium bicarbonate was added, and the pH was adjusted to 9. Acid. the mixture was left to pH 1.0 with dilute HCl and extracted with chloroform. The extract was dried over MgSO4, filtered and evaporated to dryness to give the desired compound as an oil.

(b) Methyl 4 - [(2-carbomethoxyethyl) thio] -4-carboxybutanoate

19tb)

To a stirred solution of 29.0 g (0.24 M) of methyl 3-mercaptopropionate and 61.0 g (0.60 M) of triethylamine in 200 ml of methanol was added 22.5 ml (0.10 M) of 19 (a) under an argon atmosphere. After stirring at room temperature for 10 minutes, the reaction was refluxed for 1.5 hours, cooled to room temperature and concentrated in vacuo. The residue was then diluted with ethyl acetate, extracted with 5% sodium bicarbonate, acidified to pH 1.6 with dilute HCl and extracted several times with chloroform

SKB CASE 14135-58-59

JjBBlF * "

- 38 -

zes. The organic extracts were then combined, dried over MgSO4, filtered and evaporated to dryness to give the desired compound.

(c) methyl 4- [2- (carbomethoxyethyl) thio] -5-hydroxypentanoate 19 (c)

A solution of 19 (b) (21.4 g, 0.08 M) in THF (175 ml) was cooled to-160 ° C in a methanol-ice bath. The contents were allowed to cool for 10 minutes, at which time borane-methyl sulfide complex (7.2 g, 0.09 M) was added slowly. After the addition the reaction was warmed to room temperature β then refluxed for about 15 minutes. After cooling, acetic acid (6 ml) was added and excess THF was evaporated. The reaction mixture was dissolved in ethyl acetate, washed with 5% sodium bicarbonate, dried over MgSO4, filtered, evaporated to dryness and the crude alcohol was obtained. This material was then instantly chromatographed employing a 60% ethyl acetate in hexane system to give a purest product.

(d) methyl 4- [2- (carbomethoxyethyl) thio] -4-formylbutanoate

19 (d)

Methylene chloride (20 ml) and Me2 SO (1.14 ml, 0.016 M) were combined and cooled to -65Â ° C. TFAA (0.91 mL, 0.012 M) was added dropwise to the cold solution, at which point a precipitate formed. Five minutes later, a sol. (2.0 g, 0.008 M) in methylene chloride (3 ml), keeping the temperature at -65 ° C. The reaction was then stirred for 30 minutes, followed by the addition of TEA (3.1 mL, 0.022 M) dropwise. The reaction mixture was stirred for 30 minutes at -65 ° C, at which point it was diluted with methylene chloride, washed with dilute HCl, water and then twice with 5/6 sodium bicarbonate. The organic phase was then dried over MgSO4, filtered and evaporated to dryness to give the crude aldehyde. This material was flash chromatographed employing 50.6 g of ethyl acetate in hexane to give a more pure product.

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(and)

-39-

Methyl 4- (2-carbomethoxyethyl) thio] -5 (Z) -octadecenoate

19iâi

A mixture of tridecyltriphenylphosphonium bromide (1.27 g, 2.4 mmol) and THF (20 mL) was stirred at ambient temperature for 3 minutes. The solution was then cooled to -68Â ° C, where n-butyllithium (1.05 mL, 2.4 mmol) was added slowly maintaining the reaction temperature below -65Â ° C. After stirring at -68Â ° C the reaction was allowed to warm to -1Â ° C for about 10 minutes. After recooled to -89 ° C, 19 (d) (0.05 g, 2.0 mmol) was dissolved in THF (10 mL) and slowly added. The reaction was stirred for 1 hour at -68 ° C, then the bath was removed. The product was then triturated to hexane as well as ether, the hexane and ether fractions combined, evaporated to dryness and the residue remaining flash chromatographed to give the desired octadecenoate, (f) 4- (2-carboxyethyl) thio] -5 (Z) -octanoic acid

19 (e) (319 mg, 0.826 mmol) were dissolved in methanol (7.5 mL) and stirred under argon at &lt; 0 &gt; C. An IN solution of sodium hydroxide (3.1 mL, 3.08 mmol) was added dropwise. The ice bath was then removed and the reaction was allowed to warm to room temperature for about 3 hours, then refrigerated overnight. Most of the methanol was then evaporated and the aqueous residue cooled in an ice bath, then acidified with dilute HCl. The aqueous phase was extracted twice with diethyl ether. The ether extracts were then combined and dried over magnesium sulfate, filtered, and evaporated to give crude product. This was recrystallized from diethyl ether-hexane and the final product was obtained, mp. Comp. calculated: C 65.24; H 9.91; S, 8.29. Found: C 65.50; H, 10.05; S, 8.61.

EXAMPLE 20

As a specific example of a composition of this invention,

an active ingredient, such as the compound of Example 4 (i), is

dissolved in sterile water at a concentration of 0.5% and converted into aerosol from a vaporizer with an air flow

62 368

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frog

To provide the desired aerosol drug weight EXAMPLE 21

As a further specific example of a composition of this invention, an active ingredient, such as the compound of Example 5, is mixed with mannitol at a concentration of 1.0% and administered from a regulated powder inhalation device to provide the weight of intended drug.

Claims (8)

A process for the preparation of the compounds represented by the formula (I) S (CH ₂ ) _m CHCOR ₂ ^CH 3 ^(CH 2 ⁾ p Z ^CH = ^X c = c (I) wherein m is 0, 1 or 2; p is 9, 10, 11, 12 or 13; FL 'is hydrogen, fl amino or -NHCCH3; R ₂ is hydroxy, amino, -NHCH ₂ CO ₂ H, -NCH ₂ CO ₂ H, CH ₃ -NHCHCO ₂ H, -NHCHCO ₂ H or NHCH _{2 C 2} H; and X is -C 0 -C ₂ H, -CH ₂ 0H, CHch (ch ₃ ) ₂ -CH ₂ OH or -CH (CH ₂ ) _n COO, where n is 1 or 2, Y is hydrogen or H and Y and R is hydroxy or amino, provided that when m is 0, R1 is hydrogen, or a respective pharmaceutically acceptable salt thereof, comprising: (A) when X is -CH (CH ₂ ) ₃ COR, where Y is hydroxyl and n is 2, the reaction of (1) CH ₂ (CH ₂ ) _p C = C 0 / \ CH ^ CHCH ₂ CH ₂ C0r _3, where R is R H 2 ^X H fl ' or a readily convertible radical in R, with (2) HS (CH ₂ ) _n CHCOR ₂ ' 62 368 SKB CASE 14135-58-59 -41- where R '8 R ° ^s ^ ₂ * R ^ respectively and R radical or an facilmer' convertible to R ₂ and R ₂ ; (8) when X is -H (CH ₂ ) _n CQR, where Y is hydroxy in 6, YR, S (CH ₂ ) m CHCOR ₂ the reaction of (3) CH, (CH 2) CHCH / ^C Z Η H with (4) CHâ,ƒCNRâ,ƒ; (C) when X is -CH (CH ₂ ) _n CQR, where Y is hydrogen, the reacS (CH-) CHCOR ' (5) HCCHCH ₂ (CH ₂ ) _n CO ₃ with (O) CH 2 Cl 3) CH = P 2; (D) when X is -CCLH, the reaction of (7) (CH) CHCOR '' 2 m 2 ^CH 3 ^(CH 2 ⁾ p CHCHQ with (8) an oxidizing agent; c = c (E) when X is -CH "OH, the reaction of (7) fi ' (CHâ,,) with (9) a reducing agent; or (F) when X is -CHCH2 QH, the reaction of (10) OH ch ₃ (ch ₂ ) _Px C = C Z i CH-CHCH ₂₀ OH with (2) HS (CH ₂ ) CHCOR 4 ''; ch ₃ (ch ₂ ) ^ chcho "" c = c ^ H ^X H 62 368 SKB CASE 14135-58-59 4 And thereafter the conversion of R ₂ , R ₂ 'to R ₂ into R ₂ , R ₂ and R 5 at the option is the formation of the pharmaceutically acceptable salt of the compounds. A process according to claim 1 for the preparation of the compounds of formula III ^CH 3 ( ^CH 2) S (CH ₂ ) _ro CHCOR ₂ CH ₂ H (CH ₂ ) _n COO C = CY (III) wherein n is 2 and Y is hydroxy, comprising the reaction of THE (1) CH (CH ₂₎ CHCH ₂ CH _SM-2 with C0Rj X Η H ^R 1 ' I (2) HS (CH ₂ ) _m CHCOR ₂ '. A process according to claim 2 for the preparation of 5 - [(2-amino-3-carboxymethylamino-3-oxopropyl) thio] -4-hydroxy-6 (Z) -nonadecenoic acid, comprising reacting 4,5-epoxy-6 (Z) -nonadecenoate with 2-trifluoroacetamido-3-carbomethoxymethylamino-3-oxopropylmercaptan and the conversion of the resulting aqueous base compound to the desired product. A process according to claim 2 for the preparation of 4-hydroxy-5- (2-carboxyethyl) thio] ^- (Z) -nonadecenoic acid, comprising reacting 4,5-epoxy-6- (Z) methyl-3-mercaptopropionate and the conversion of the resulting aqueous base compound to the desired product. A process according to claim 1 for the preparation of the compounds of formula III ^CH 3 ^CH 2 ⁾ p. , / c = c S (CH) _The CHCQR | 2 m 2 , CHIPH (CH ₂ ) _n C0R Y (III) H 62 368 SKB CASE 14135-58-59 r Wherein n is 6 and Y is hydroxy, comprising the reaction of fl ' (3) ^CH3 CH2 ^CH2 / H CHCOR 'm 2 CCHO H with (4) CH ₃ CO ₃ . A process according to claim 5 for the preparation of 3-hydroxy-4 - [(2-carboxyethyl) thio] -5 (Z) -octadecenoic acid, comprising reacting 2 - [(2-carboxyethyl) thiohexadec-3 (Z) -enenal with methyl acetate and conversion of the resulting aqueous base compound to the desired product. Process according to claim 1 for the preparation of the compounds of formula III I ¹ S (CH) CHC0R _the | 2 m 2 CH (CH) CHCH (CHâ) COR (ILI) 3 "2'p χ | 2 n C = C Y Η H in which Y is hydrogen, comprising the reaction of (5) I ¹ S (CH) CHCOR. · I '2'ra 2 HCCHCH "(CH) _x Color with (6) CH (CH) CH = P ^. || ZZn>> Z p 0 8. A process according to claim 7 for the preparation of 4- [2- (carboxyethyl) thio] -5 (Z) -octadecenoic acid, which comprises reacting methyl 4- [2- (2-carbomethoxyethyl) -4-formylbutanoate with tridecyltriphenylphosphonium ylide and conversion of the resulting compound to aqueous base in the desired product.